A growing subset of head and neck cancers are caused by infection with the human papillomavirus (HPV) type-16. There is a strong need to develop novel treatment strategies which take advantage of the unique tumor biology of this patient cohort. HPV-specific vaccines represent such a promising, targeted adjuvant treatment option. We plan to evaluate a DNA vaccine, pNGVL4a-CRT/E7(detox) DNA, in combination with various strategies that can further enhance its potency. In addition to evaluating the safety of the novel DNA vaccine, we plan to measure immunologic responses and characterize tumor infiltrating immune cells within pathologic specimens which will be obtained before and after DNA vaccination as part of routine standard of care. This will give us an opportunity to better understand how this environment may change after vaccination, which will be critical to successful immunotherapy translation. The results of this phase I clinical trial will facilitate a subsequent phase II clinical trial which will evaluate the efficacy of the vaccine in generating HPV-specific CD8+ T cell immune responses which can potentially impact the survival in this patient population by providing long-term immune protection against the development of locoregional recurrence and/or micrometastatic disease. The strengths of our application include the availability of GMP grade reagents developed by the research team and NCI RAID, and previous experience in DNA vaccine clinical trial design and execution with an infrastructure in place at our institution which facilitates further investigation into this area. In addition, we have support from the institution to conduct this clinical trial and access to supportive collaborators who are also evaluating human immune responses elicited after vaccination. The project challenges existing paradigms by coupling chemoradiation treatment with immunotherapy in order to achieve a more potent immunologic response. The results gained from this study will advance the field of immunotherapy by providing information which will allow us to re-evaluate our current strategies to enhance vaccine potency.